Novel nucleic acid molecules and their use in therapy

Abstract

Disclosed are products and methods for therapy using nucleic acid molecules, and in particular in relation to treatment of alpha-1-antitrypsin deficiency. Also disclosed are pharmaceutical compositions including the nucleic acids and/or delivery vehicles including the nucleic acids, and their use in manufacture of pharmaceutical compositions for use in therapy, such as treatment of alpha-1-antitrypsin deficiency.

1 . A chemically modified ribonucleic acid consisting of 19, 20, or 21 nucleotides and having a sequence according to SEQ ID NO: 39, and 40. 
     
     
         2 . The ribonucleic acid according to  claim 1 , wherein the chemically modified ribonucleic acid is modified in the phosphodiester backbone, or in the sugar backbone. 
     
     
         3 . The ribonucleic acid according to  claim 1 , wherein the chemically modified ribonucleic acid is modified in the phosphodiester backbone by incorporation of phosphorothioate, boranophosphate, or methylphosphonate. 
     
     
         4 . The ribonucleic acid according to  claim 1 , wherein the chemically modified ribonucleic acid is modified in the sugar backbone at the 2′-position of the ribose unit, by substitution of the 2′-OH group for —O—CH 3 , —CH 2 CH 2 OCH 3 , or —F, or by incorporation of 2-thiouridine, 5-methylcytidine or pseudouridine. 
     
     
         5 . An RNA molecule consisting of 46-100 nucleotides, comprising two sequences spaced 4-10 nucleotides apart, wherein the two sequences are selected from the sequence pairs: SEQ ID NO: 39 and 40. 
     
     
         6 . A DNA molecule comprising at least one nucleotide sequence complementary to the RNA molecule according to  claim 1 , wherein the nucleotide sequence is operably linked to a single RNA polymerase promoter sequence. 
     
     
         7 . A DNA molecule comprising at least two nucleotide sequences, each complementary to the RNA molecule according to  claim 1 , wherein each nucleotide sequence is independently operably linked to an RNA polymerase promoter sequence and wherein the RNA polymerase promoter sequence is the same for each nucleotide sequence. 
     
     
         8 . A DNA molecule comprising at least two nucleotide sequences, each complementary to the RNA molecule according to  claim 1 , wherein each nucleotide sequence is independently operably linked to an RNA polymerase promoter sequence and wherein the RNA polymerase promoter sequence is different for each nucleotide sequence. 
     
     
         9 . The DNA molecule according to  claim 6 , wherein each RNA polymerase promoter sequence is selected from RNA polymerase promoters H1, 7SK, and U1. 
     
     
         10 . A virus particle comprising a recombinant viral genome, wherein said genome comprises a DNA molecule according to  claim 6 . 
     
     
         11 . A virus particle comprising a recombinant viral genome, wherein said genome comprises an RNA molecule comprising a nucleotide sequence complementary to a DNA molecule comprising at least one nucleotide sequence complementary to at least one RNA molecule consisting of 46-100 nucleotides, comprising two sequences spaced 4-10 nucleotides apart, wherein the two sequences consist of the sequence pair: SEQ ID NO: 39 and 40. 
     
     
         12 . A vehicle selected from the group plasmid DNA, lipid-based vectors, and polymeric vectors, and comprising the DNA molecule according to  claim 6 . 
     
     
         13 . A method for treatment of alpha-1-antitrypsin deficiency, said method comprising administering a nucleic acid molecule selected from the group consisting of chemically modified ribonucleic acid according to  claim 1 , a ribonucleic acid molecule consisting of 21 nucleotides and having a sequence selected from SEQ ID NO: 39 and 40, and a nucleic acid molecule consisting of 46-100 nucleotides, comprising two sequences spaced 4-10 nucleotides apart, wherein the two sequences consist of the sequence pair: SEQ ID NO: 39 and 40, to a subject in need thereof. 
     
     
         14 . The method for treatment according to  claim 13 , wherein the nucleic acid molecule is administered using a vehicle delivered to the subject by way of gene gun/ballistic DNA, electroporation, sonoporation, hydroporation, magnetofection, needle injection and/or other methods facilitating the incorporation of DNA or RNA into the cell leading to a modified transcription and expression of target genes, wherein the vehicle is for delivery of nucleic acid material to a human cell in vivo. 
     
     
         15 . The method for treatment according to  claim 13 , wherein the subject's genome is heterozygous or homozygous for a G342K mutation in a gene encoding alpha-1-antitrypsin. 
     
     
         16 . The method for treatment according to  claim 13 , wherein the alpha-1-antitrypsin deficiency manifests as liver cirrhosis, pulmonary emphysema, necrotising panniculitis, systemic vasculitis, (intracranial) aneurysms, fibromuscular dysplasia, bleeding disorders, anterior uveitis, systemic necrotizing vasculitis and Wegener granulomatosis.